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Application of Rice rel1 Gene in Improving Plant Resistance to Drought Stress

A technology of drought resistance and rice, applied in the fields of application, plant products, genetic engineering, etc., to achieve the effect of increasing expression, improving resistance, and favorable leaf angle

Active Publication Date: 2020-03-17
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reason for the curling of leaves caused by REL1 is the increase of vesicular cells. Therefore, whether the REL1 gene has other functions or applications is still unknown

Method used

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  • Application of Rice rel1 Gene in Improving Plant Resistance to Drought Stress
  • Application of Rice rel1 Gene in Improving Plant Resistance to Drought Stress
  • Application of Rice rel1 Gene in Improving Plant Resistance to Drought Stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 PEG handles blade curling degree

[0032] The typical manifestation of rice water loss under drought stress is leaf involution. To identify the drought resistance of wild-type (WT), rel1 mutant and overexpressed REL1 (OE3) rice, the results of different concentrations of PEG (polyethylene glycol) treatment The leaves are observed, the specific steps are as follows:

[0033] (1) Wild-type, rel1 mutant and overexpressed OE3 rice were germinated at 37°C, then transferred to 1 / 2 MS medium, and cultured at 28°C for 14 days under 16 hours of light (natural light) and 8 hours of darkness.

[0034] (2) Treat the plants obtained in step (1) with 0%, 10%, 20%, 30%, 40% (w / w) PEG6000 solutions for 24 hours respectively.

[0035] (3) Observe the cross-section of the leaves of the treated plants and analyze the results. The result is as figure 1 As shown, under 0% PEG treatment, both the rel1 mutant and the overexpressed OE3 showed outward curling of leaves, while WT...

Embodiment 2

[0036] Embodiment 2SOD activity assay

[0037] SOD activity is considered to be an important indicator of drought resistance. In order to identify the drought resistance of wild-type, rel1 mutant and overexpressed REL1 (OE3) rice, the SOD activity after PEG treatment was determined, and the specific steps were as follows:

[0038] (1) The cultivation method of paddy rice is the same as embodiment 1.

[0039] (2) The plants obtained in step (1) were treated with 20% (w / w) PEG6000 solution, and the SOD activity was measured 3 hours, 6 hours, 12 hours and 24 hours after the treatment.

[0040] (3) Analysis of SOD activity assay results. The result is as figure 2 As shown, the SOD activity of wild type, rel1 mutant and overexpressed OE3 all decreased at 6 hours after treatment and peaked at 12 hours. In the three processes of 3-6 hours, 6-12 hours and 12-24 hours, the order of increase and decrease of SOD enzyme activity was as follows: rel1>OE3>WT.

Embodiment 3

[0041] Example 3 Differences in Expression of Drought Resistance Genes between Wild Type and rel1 Mutants

[0042] In order to find the molecular evidence of the drought-resistant phenotype of the rel1 mutant, the expression levels of five drought-resistant genes (LOC_Os05g12400, LOC_Os05g12640, LOC_Os10g26940, LOC_Os11g06980, LOC_Os03g20090) in the wild type and rel1 mutant were detected. Specific steps are as follows:

[0043] (1) RNA of wild type and rel1 mutant was extracted by Trizol method.

[0044] (2) Use TransScript First-Strand cDNA Synthesis SuperMix of TransGen Biotech Company to perform reverse transcription first-strand (cDNA) synthesis, and operate according to the instructions.

[0045] (3) Use Takara's Premix Ex Taq TM II (Tli RNaseH Plus) kit is used for fluorescent quantitative PCR detection of the cDNA synthesized in step (2), wherein the primer pairs are respectively Primer1 / 2, Primer3 / 4, Primer5 / 6, Primer7 / 8, Primer9 / 10, internal reference The primer p...

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Abstract

The invention discloses application of a rice REL1 gene to the improvement of drought stress resistance of plants. Moderate lamina warping and reasonable leaf include angle are favorable for survivaland development of rice under the stress condition. In order to expand the application of the rice REL1 gene to rice drought stress resistance, wild type, rel1 mutant and overexpression REL1 rice materials are subjected to drought stress treatment, lamina inward warping and SOD activity variable amplitude serve as monitoring indexes, and the invention provides that the drought stress resistant ability of the rel1 mutant and the overexpression REL1 rice is improved relative to that of the wild type; meanwhile, 5 drought-resistant genes are subjected to expression analysis simultaneously, and the invention proves that upregulation of the REL1 gene expression quantity can improve the drought stress resistance of the rice physiologically and molecularly. The drought stress resistance of the rice is enhanced by increasing the gene expression quantity of the REL1, and a new choice is provided for genetic improvement and molecular breeding of rice drought resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of rice REL1 gene in improving plant resistance to drought stress. Background technique [0002] Leaf morphology affects the photosynthetic capacity and transpiration of rice, and is an important part of the ideal plant architecture. Plant leaves are usually polarized into a proximal-distal end, so under adverse conditions two different leaf morphologies develop: abaxially curled and adaxially curled. [0003] The rice rel1 mutant exhibited curled leaves, increased leaf angles, and decreased plant height and yield. Electron microscopy revealed that the rel1 phenotype was due to increased size and number of vesicular cells. The REL1 gene consists of two exons and one intron, encodes an unknown protein, and is highly conserved in monocots. Knocking down the expression of REL1 in the rel1 mutant can restore the phenotype, while up-regulating the expression...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
Inventor 张泽民谢庆军梁嘉燕陈雄辉彭海峰孙博
Owner SOUTH CHINA AGRI UNIV
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